Compositions and methods having improved efficacy against spores and other organisms

ABSTRACT

Compositions and methods for the disinfection of surfaces are provided. The compositions include an alcohol and a peracid. The disinfectant composition is characterized by a pH of no more than about 5. Broad spectrum efficacy is achieved, and synergistic activity is exhibited against bacterial and fungal spores, including  Clostridium difficile  spores. The compositions and methods are also useful against conformationally altered prions, bacteria, fungi, and viruses.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication Ser. No. 61/937,060, filed Feb. 7, 2014, which isincorporated herein by reference.

TECHNICAL FIELD

Embodiments of the present invention provide compositions and methodshaving surprising efficacy against bacterial and fungal spores,including Clostridium difficile spores. Embodiments are also usefulagainst conformationally altered prions, bacteria, fungi, and viruses.Embodiments of the present invention provide acidified alcoholiccompositions containing one or more peracids.

BACKGROUND OF THE INVENTION

Different types of organisms vary in their response to antiseptics anddisinfectants. The microbial susceptibility to antiseptics anddisinfectants of various organisms has been summarized as shown in thefollowing schematic. McDonnell, Gerald et al., “Antiseptics andDisinfectants: Activity, Action, and Resistance,” Clinical MicrobiologyReviews, Vol. 12, pp. 147-179 (January 1999). Organisms are listed indescending order of resistance.

-   -   Prions (CJD, BSE)

    -   

    -   Coccidia (Cryptosporidium)

    -   

    -   Spores (Bacillus, C. difficile)

    -   

    -   Mycobacteria (M. tuberculosis, M. avium)

    -   

    -   Cysts (Giardia)

    -   

    -   Small non-enveloped viruses (Polio virus)

    -   

    -   Trophozoites (Acanthamoeba)

    -   

    -   Gram-negative bacteria (non-sporulating) (Pseudomonas,        Providencia)

    -   

    -   Fungi (Candida, Aspergillus)

    -   

    -   Large non-enveloped viruses (Enteroviruses, Adenovirus)

    -   

    -   Gram-positive bacteria (S. aureus, Enterococcus)

    -   

    -   Lipid enveloped viruses (HIV, HBV)

Bacterial spores of the genera Bacillus and Clostridium have been widelystudied and are considered to be the most resistant of all types ofbacteria to antiseptics and disinfectants.

Clostridium species are significant pathogens. Infections among patientsin healthcare facilities caused by the bacteria Clostridum difficile (C.difficile), are at historically high levels. C. difficile is aspore-forming, Gram-positive anaerobic bacillus of the human intestineand is thought to be present in 2-5% of the adult population. PathogenicC. difficile strains produce multiple toxins, the mostwell-characterized of which are enterotoxin (Clostridium difficile toxinA) and cytotoxin (C. difficile toxin B), both of which can producediarrhea and inflammation in infected patients. The emergence of a new,highly toxic strain of C. difficile, resistant to flouroquinoloneantibiotics, such as ciprofloxacin and levofloxacin have also beenreported. C. difficile infection causes diarrhea and other intestinalproblems and is linked to 14,000 deaths in the United States each year.

Control of C. difficile outbreaks present significant challenges tohealth care facilities. C. difficile spores survive routineenvironmental cleaning with detergents and hand hygiene withalcohol-based gels. The spores can survive on surfaces for long periodsof time. As a result, the bacteria can be cultured from almost anysurface. Once spores are ingested, their acid-resistance allows them topass through the stomach unscathed. They germinate and multiply intovegetative cells in the colon upon exposure to bile acids.

A variety of strategies have been proposed to kill C. difficile sporeson various surfaces, with limited success. Bleach-based compositionshave been employed for hard surfaces, and have been shown to reduce theenvironmental burden of C. difficile but can be corrosive. Hydrogenperoxide-based compositions have also been proposed, includingcombinations of hydrogen peroxide and peracetic acid, a combination ofhydrogen peroxide and silver cation dry-mist system, and the so-calledAccelerated Hydrogen Peroxide (AHP). Peracids generally have poorstability, odor and corrosive properties. Hydrogen peroxide is alsoprone to decomposition, and concentrated solutions can be highlycorrosive. Alcohol-based sanitizers have not generally been effective.In fact, ethanol is sometimes used to store C. difficile spores.

A need remains for more stable, less corrosive compositions having goodefficiency against C. difficile spores, as well as against otherpathogenic infectious agents such as bacteria, fungi, viruses, fungaland bacterial spores, and conformationally altered prions, some of whichcan be particularly resistant to current sanitizers and cleansers.

Somewhat relatedly, there is growing concern about the use of biologicaltoxins and infectious agents such as conformationally altered prions,bacteria, viruses, and fungal and bacterial spores, as biological andchemical warfare agents. Chemical and biological warfare agents can befast-acting and pervasive. There is a continuing need for effective,easy to use products that will be safe for humans and the environment,that can be used to decontaminate skin, and particularly wounds,following chemical and/or biological warfare agent exposure.Furthermore, there is a need for effective products that can be used todecontaminate homes, building materials, and furniture that candecontaminate surfaces to eliminate these infectious agents, and thatcan reduce the transmission of the infectious pathogens.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic view of a dispenser according to the presentinvention.

FIG. 2 is a schematic representation of the log reduction (CFU/ml) of C.difficile spores for various test compositions, as described herein.

FIG. 3 is a schematic representation of the log reduction (CFU/ml) of C.difficile spores for various test compositions, as described herein.

FIG. 4 is a schematic representation of the log reduction (CFU/ml) of C.difficile spores for various test compositions, as described herein.

FIG. 5 is a schematic representation of the log reduction (CFU/ml) of C.difficile spores for various test compositions, as described herein.

FIG. 6 is a schematic representation of the log reduction (CFU/ml) of C.difficile spores for various test compositions, as described herein.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

In one or more embodiments, the present invention provides adisinfectant composition. The physical form of the disinfectantcomposition is not particularly limited, and in one or more embodiments,the composition may be presented as a liquid that is poured, pumped,sprayed, or otherwise dispensed, a gel, an aerosol, or a foam, includingboth aerosol and non-aerosol foams. The disinfectant composition of thepresent invention may be employed on a wide variety of surfaces orsubstrates, including hard surfaces, soft surfaces, non-living surfaces,living tissue, skin, soil, porous, and non-porous surfaces. Thecompositions of the invention may be employed to disinfect or otherwisesanitize inanimate objects such as instruments, medical and militaryequipment, furniture, handrails, textiles, etc. In one or moreembodiments, the disinfectant composition may be presented as a wipe,i.e. a tissue or cloth that is wiped over a surface.

The disinfectant composition comprises at least one alcohol. In one ormore embodiments, the alcohol is a C₁₋₆ alcohol, i.e. an alcoholcontaining 1 to 6 carbon atoms. Such alcohols may be referred to aslower alkanols. Typically, these alcohols have antimicrobial properties.Examples of lower alkanols include, but are not limited to, methanol,ethanol, propanol, butanol, pentanol, hexanol, and isomers and mixturesthereof. In one or more embodiments, the alcohol comprises ethanol,propanol, or butanol, or isomers or mixtures thereof. In one or moreembodiments, the alcohol comprises isopropanol. In other embodiments,the alcohol comprises ethanol. In one or more embodiments, thedisinfectant compositions comprise a mixture of alcohols. In one or moreembodiments, the disinfectant compositions comprise a mixture of ethanoland isopropanol. In one or more embodiments, the disinfectantcompositions comprise a mixture of isopropanol and n-propanol.

Generally, the disinfectant composition comprises at least about 10percent by weight (wt. %) alcohol, based upon the total weight of thedisinfectant composition. In one embodiment, the disinfectantcomposition comprises at least about 15 weight percent alcohol, inanother embodiment, the disinfectant composition comprises at leastabout 20 weight percent alcohol, in another embodiment, the disinfectantcomposition comprises at least about 25 weight percent alcohol, in yetanother embodiment, the disinfectant composition comprises at leastabout 30 weight percent alcohol, and in still yet another embodiment,the disinfectant composition comprises at least about 40 weight percentalcohol, based upon the total weight of disinfectant composition. In oneembodiment, the disinfectant composition comprises at least about 50weight percent alcohol, in another embodiment, the disinfectantcomposition comprises at least about 60 weight percent alcohol, inanother embodiment, the disinfectant composition comprises at leastabout 65 weight percent alcohol, in yet another embodiment, thedisinfectant composition comprises at least about 70 weight percentalcohol, and in still yet another embodiment, the disinfectantcomposition comprises at least about 78 weight percent alcohol, basedupon the total weight of disinfectant composition. More or less alcoholmay be required in certain instances, depending particularly on otheringredients and/or the amounts thereof employed in the composition. Incertain embodiments, the disinfectant composition comprises from about10 weight percent to about 98 weight percent alcohol, in otherembodiments, the disinfectant composition comprises from about 15 weightpercent to about 95 weight percent of alcohol, in yet other embodiments,the disinfectant composition comprises from about 20 weight percent toabout 90 weight percent of alcohol, and in still other embodiments, thedisinfectant composition comprises from about 30 weight percent to about85 weight percent of alcohol, based upon the total weight of thedisinfectant composition. In certain embodiments, the disinfectantcomposition comprises from about 50 weight percent to about 98 weightpercent alcohol, in other embodiments, the disinfectant compositioncomprises from about 60 weight percent to about 95 weight percent ofalcohol, in yet other embodiments, the disinfectant compositioncomprises from about 65 weight percent to about 90 weight percent ofalcohol, and in still other embodiments, the disinfectant compositioncomprises from about 70 weight percent to about 85 weight percent ofalcohol, based upon the total weight of the disinfectant composition.

In any of the above embodiments, the disinfectant composition mayfurther include a peracid. Examples of peracids include C₁-C₁₂percarboxylic acids, diperoxyglutaric acid, diperoxyadipic acid,diperoxysuccinic acid, diperoxysuberic acid, diperoxymalonic acid,peroxylactic acid, peroxyglycolic acid, peroxyoxalic acid, peroxypyruvicacid (PPA), and mixtures thereof. Examples of peracids also includeperoxyoctanoic acid. In one or more embodiments, the peracid is selectedfrom C₃₋₁₂ peracids. In one or more embodiments, the peracid is selectedfrom C₃₋₁₀ peracids. In one or more embodiments, the disinfectingcompositions include a mixture of peracids. In one or more embodiments,the peracid comprises peroxypyruvic acid. In one or more embodiments,the disinfecting compositions include a peracid other than peraceticacid. In other embodiments, the disinfecting compostions include aperacid in addition to peracetic acid. See U.S. Pat. Nos. 4,051,058,4,051,059, 5,200,189, 5,200,198, 5,489,434, 5,718,910, 5,314,687,5,437,868, and U.S. Pub. Pat. App. Nos. 2010/0261792 A1 and 2012/0213835A1 for further discussion on peracids. These patents are incorporatedherein by reference in their entirety.

In one embodiment, the peracid is added to the disinfectant compositionas a solution or emulsion. In other words, the peracid may be premixedwith a carrier, and optionally one or more other ingredients, to form aperacid solution or emulsion, with the proviso that the carrier does notdeleteriously affect the sporicidal properties of the composition. Morespecifically, a carrier deleteriously affects the sporicidal propertiesof the composition when it decreases the log kill by more than a deminimus amount. By de minimus is meant a decrease of less than about 0.5log kill.

Examples of carriers include water, alcohol, or blends of water andanother carrier such as glycols, ketones, linear and/or cyclichydrocarbons, triglycerides, carbonates, silicones, alkenes, esters suchas acetates, benzoates, fatty esters, glyceryl esters, ethers, amides,polyethylene glycols, PEG/PPG copolymers, inorganic salt solutions suchas saline, and mixtures thereof. It will be understood that, when theperacid is premixed to form a peracid solution or emulsion, the amountof solution or emulsion that is added to the disinfectant composition isselected so that the amount of peracid falls within the ranges set forthhereinabove.

Advantageously, a synergistic antimicrobial and/or sporicidal effect isobserved when the peracid is combined with alcohol at an acidic pH. Incertain embodiments, peracids that exhibit little or no efficacy ontheir own against C. difficile spores or other resistant organismsprovide an enhanced efficacy when combined with alcohol according to thepresent invention, and a further enhanced efficacy when the pH of thedisinfectant composition is less than 7. It has surprisingly been foundthat, while disinfectant compositions show little or no efficacy againstsome bacterial or fungal spores, the combination of an enhancer andalcohol at a low pH exhibits synergistically enhanced efficacy againstthe spores, such C. difficile spores. Also, it has been unexpectedlyfound that the methods of the present invention provide good efficacyagainst C. difficile spores at much lower concentrations of peracid thanwhat is generally thought to be required for good efficacy.

In one or more embodiments, the amount of peracid is 2 weight percent(wt. %) or less, in other embodiments, 1.5 wt. % or less, in otherembodiments, 1 wt. % or less, in other embodiments, 0.9 wt. % or less,based upon the total weight of the disinfectant composition.

In one or more embodiments, the peracid may be present in an amount ofat least about 100 parts per million by weight (ppm), based upon thetotal weight of the disinfectant composition, in other embodiments, atleast about 200 ppm, in other embodiments, at least about 500 ppm, inother embodiments, at least about 1000 ppm, based upon the total weightof the disinfectant composition.

In one or more embodiments, the peracid may be present in an amount offrom about about 100 ppm to about 2 weight percent (wt. %), based uponthe total weight of the disinfectant composition. In other embodiments,the amount of peracid is from about 200 ppm to about 1.5 wt. %, and inyet other embodiments, from about 500 ppm to about 1 wt. %, based uponthe total weight of the disinfectant composition. These amounts, whichare lower than what is found in previous peracetic acid-based productsand other peracid-based products, are safer for use on skin. Forexample, many previous surface disinfectants that are based upon acombination of peracetic acid and bleach recommend that the user weargloves while using the product to avoid skin contact.

In one or more of the above embodiments, the pH of the disinfectantcomposition is less than about 5; in one or more embodiments, the pH ofthe disinfectant composition is less than about 4.5; in one or moreembodiments, the pH of the disinfectant composition is less than about4; in one or more embodiments, the pH of the disinfectant composition isless than about 3.5; in one or more embodiments, the pH of thedisinfectant composition is less than about 3; in one or moreembodiments, the pH of the disinfectant composition is less than about2.5; in one or more embodiments, the pH of the disinfectant compositionis less than about 2.

In one or more of the above embodiments, the pH of the disinfectantcomposition is from about 0 to about 5. In one or more of the aboveembodiments, the pH of the disinfectant composition from about 0.5 toabout 3. In one or more of the above embodiments, the pH of thedisinfectant composition from about 1 to about 2. The disinfectantcomposition may therefore be referred to as acidified, since thedisinfectant composition has an acidic pH.

The disinfectant composition may be acidified by the addition of one ormore acids. The type of acid is not limited, however, weak acids are notpreferred. The acid should have a pKa of 5.4 (the pKa of citric acid) orless.

Examples of useful acids include mineral acids and organic acids.Mineral acids include, without limitation, hydrochloric acid, nitricacid, phosphoric acid, phosphonic acid, boric acid, and sulfuric acid.Organic acids include sulfonic acids, organophosphorus acids, carboxylicacids such as benzoic acids, propionic acids, phthalic acids, butyricacids, acetic acids, amino acids, and other substituted andunsubstituted organic acids.

Examples of organic acids include adipic acid, benzene 1,3,5tricarboxylic acid, chlorosuccinic acid, choline chloride, cis-aconiticacid, citramalic acid, citric acid, cyclobutane 1,1,3,3 tetracarboxylicacid, cyclohexane 1,2,4,5 tetracarboxylic acid, cyclopentane 1,2,3,4tetracarboxylic acid, diglycolic acid, fumaric acid, glutamic acid,glutaric acid, glyoxylic acid, isocitric acid, ketomalonic acid, lacticacid, maleic acid, malic acid, malonic acid, nitrilotriacetic acid,oxalacetic acid, oxalic acid, phytic acid, p-toluenesulfonic acid,salicylic acid, succinic acid, tartaric acid, tartronic acid,tetrahydrofuran 2,3,4,5 tetracarboxylic acid, tricarballylic acid,versene acids, 3-hydroxyglutaric acid, 2-hydroxypropane 1,3 dicarboxylicacid, glyceric acid, furan 2,5 dicarboxylic acid, 3,4-dihydroxyfuran-2,5dicarboxylic acid, 3,4-dihydroxytetrahydrofuran-2,5-dicarboxylic acid,2-oxo-glutaric acid, dl-glyceric acid, and 2,5 furandicarboxylic acid.

It has been found that acidifying the disinfectant composition enhancesthe efficacy of the alcoholic solutions against one or moremicroorganisms. In one or more embodiments, acidifying the disinfectantcomposition enhances the efficacy of the alcoholic solutions against oneor more types of bacterial or fungal spores. In one or more embodiments,acidifying the disinfectant composition enhances the efficacy of thealcoholic solutions against C. difficile.

The composition can further comprise a wide range of optionalingredients, with the proviso that they do not deleteriously affect thesanitizing efficacy of the composition. By deleterious is meant that thedecrease in the log reduction is not de minimus, or in other words, thelog reduction of C. difficile spores does not decrease by more thanabout 0.5. The CTFA International Cosmetic Ingredient Dictionary andHandbook, Eleventh Edition 2005, and the 2004 CTFA International Buyer'sGuide, both of which are incorporated by reference herein in theirentirety, describe a wide variety of non-limiting cosmetic andpharmaceutical ingredients commonly used in the skin care industry, thatare suitable for use in the compositions of the present invention.Nonlimiting examples of functional classes of ingredients are describedat page 537 of this reference. Examples of these functional classesinclude: abrasives, anti-acne agents, anticaking agents, antioxidants,binders, biological additives, bulking agents, chelating agents,chemical additives; colorants, cosmetic astringents, cosmetic biocides,denaturants, drug astringents, emollients, emulsifiers, externalanalgesics, film formers, foam surfactants, fragrance components,humectants, opacifying agents, plasticizers, preservatives (sometimesreferred to as antimicrobials), propellants, reducing agents, skinbleaching agents, skin-conditioning agents (emollient, miscellaneous,and occlusive), skin protectants, solvents, surfactants, foam boosters,hydrotropes, solubilizing agents, suspending agents (nonsurfactant),sunscreen agents, ultraviolet light absorbers, detackifiers, andviscosity increasing agents (aqueous and nonaqueous). Examples of otherfunctional classes of materials useful herein that are well known to oneof ordinary skill in the art include solubilizing agents, sequestrants,keratolytics, topical active ingredients, and the like.

It has been discovered that the combination of alcohol and peracidexhibits enhanced antimicrobial efficacy. Advantageously, auxiliaryantimicrobials, some of which can be harsh on skin, are not required. Incertain embodiments, the disinfectant composition does not contain anyauxiliary antimicrobial ingredients. Any antimicrobial ingredient otherthan the combination of alcohol, peracid and acid may be referred to asan auxiliary antimicrobial agent. In one embodiment, the amount ofauxiliary antimicrobial agent (including preservatives) is less thanabout 0.1 wt. %, in another embodiment, less than about 0.05 wt. %,based upon the total weight of the disinfectant composition. In anotherembodiment, the disinfectant composition is devoid of auxiliaryantimicrobial agents.

It is envisioned that, in other embodiments, auxiliary antimicrobialagents could be included, with the proviso that the antimicrobialingredient does not deleteriously affect the sanitizing properties ofthe composition. Examples of auxiliary antimicrobial agents include, butare not limited to, triclosan, also known as5-chloro-2(2,4-dichlorophenoxy) phenol (PCMX) and available fromCiba-Geigy Corporation under the tradename IRGASAN®; chloroxylenol, alsoknown as 4-chloro-3,5-xylenol, available from Nipa Laboratories, Inc.under the tradenames NIPACIDE® MX or PX; hexetidine, also known as5-amino-1,3-bis(2-ethylhexyl)-5-methyl-hexahydropyrimidine; ADBAC, alsoknown as alkyldiethylbenzylammonium chloride, or mixtures of ADBAC withother quat compounds, chlorhexidine salts including chlorhexidinegluconate and the salts ofN,N″-Bis(4-chlorophenyl)-3,12-diimino-2,4,11,14-tetraazatetradecanediimidiamide; dodecyl dimethyl ammonium chloride (DDAC);2-bromo-2-nitropropane-1; 3-diol, benzalkonium chloride; cetylpyridiniumchloride; alkylbenzyldimethylammonium chlorides; iodine; phenol,bisphenol, diphenyl ether, phenol derivatives, povidone-iodine includingpolyvinylpyrrolidinone-iodine; parabens; hydantoins and derivativesthereof, including 2,4-imidazolidinedione and derivatives of2,4-imidazolidinedione as well as dimethylol-5,5-dimethylhydantoin (alsoknown as DMDM hydantoin or glydant); phenoxyethanol; cis isomer of1-(3-chloroallyl)-3,5,6-triaza-1-azoniaadamantane chloride, also knownas quaternium-15 and available from Dow Chemical Company under thetradename DOWCIL™ 2000; diazolidinyl urea; benzethonium chloride;methylbenzethonium chloride; glyceryl laurate, transition metalcompounds such as silver, copper, magnesium, zinc compounds, hydrogenperoxide, chlorine dioxide, anilides, bisguanidines, tropolone, C₆₋₂₀diols, including C₆₋₁₂-alkane diols such as hexanediol, octanediol, anddecanediol, glyceryl caprylate/caprate (GCC), and mixtures thereof. Inone or more embodiments, the auxiliary antimicrobial agent(s) arepresent in an amount of from about 0 to about 2 wt. %, in otherembodiments, from about 0.005 to about 1.5 wt. %, in other embodiments,from about 0.1 to about 1 wt. %, based upon the total weight of thedisinfectant composition.

Advantageously, certain ingredients that have been designated ascritical to current sporicidal compositions can be limited in thedisinfectant composition of the present invention. For example,hypochlorous acid and precursors thereof are not necessary, and can belimited, if desired, to less than about 0.5 wt. %, or in anotherembodiment to less than about 0.1 wt. %, based upon the total weight ofthe disinfectant composition. In another embodiment, the disinfectantcomposition is devoid of hypochlorous acid.

In one or more embodiments, the compositions of the present inventionmay further include peracetic acid. However, in one or more embodiments,the peracid described hereinabove is not peracetic acid. In fact, thedisinfectant composition may be devoid of peracetic acid or the amountof peracetic acid may be limited to less than about 0.5 wt. %, in otherembodiments, to less than about 0.1 wt. %, based upon the total weightof the disinfectant composition.

Indeed, any component other than the alcohol and peracid is notnecessary to achieve antimicrobial efficacy and can optionally belimited to less than about 0.5 wt. %, if desired to less than about 0.1wt. %, if desired to less than about 0.01 wt. %, or if desired to lessthan about 0.001 wt. %. It will be understood that the balance of thedisinfectant composition may, in certain embodiments, include water orother suitable solvent. In one embodiment, the disinfectant compositionis devoid of any component other than alcohol, peracid, acidifier, andoptionally water or other suitable solvent.

The disinfectant composition may be prepared by simply mixing thecomponents together. In one embodiment, where one or more components isobtained as a solid powder, the disinfectant composition is prepared bya method comprising dispersing the solid powder in water or alcohol withslow to moderate agitation, and then adding other ingredients asdesired, and mixing until the mixture is homogeneous.

Advantageously, it has been found that compositions according to thepresent invention have efficacy against a broad spectrum of grampositive and gram negative bacteria, fungi, parasites, fungal andbacterial spores, enveloped and non-enveloped viruses, and prions (CJD,CWD, BSE, Scrapie). One or more embodiments of the present inventionexhibit efficacy against one or more of spores of Bacillus anthracis,Bacillus cereus, Clostridium difficile, Clostridium botulinum, andClostridium tetani.

Unexpectedly, when a peracid is combined with alcohol at a low pH,according to the present invention, sporicidal activity is enhanced,i.e. potentiated. In one or more embodiments, the disinfectantcomposition is effective at killing C. difficile spores. In one or moreembodiments, the disinfectant composition is also effective in killinggram negative and gram positive bacteria, fungi, parasites,non-enveloped and enveloped viruses. In one or more embodiments, thedisinfectant composition has rapid antimicrobial efficacy againstbacteria such as Staphylococcus aureus, methicillin-resistant S. aureus,Escherichia coli, Pseudomonas aeruginosa, Serratia marcescens, and fungisuch as Candida albicans and Aspergillus niger, and black mold sporesStachybotrys chartanim. In one or more embodiments, the disinfectantcomposition has rapid efficacy against skin microflora, includingresident and transient skin microflora.

Thus, the present invention further provides a method for killing orinactivating microbes on a surface comprising applying, to the surface,an effective amount of a disinfectant composition as described herein.Advantageously, the disinfectant composition of the present inventionmay be used as a healthcare personnel hand wash. In one or moreembodiments, the present invention provides an disinfectant compositionthat meets the standards of the FDA Tentative Final Monograph forHealthcare Antiseptic Drug Products (TFM) (Federal Register 59 [116],Jun. 17, 1994: pp. 31402-31452) for healthcare personnel hand wash,which is incorporated by reference herein.

The disinfectant composition and method of the present inventionprovides rapid antimicrobial efficacy upon a single use, withoutrequiring auxiliary antimicrobial agents. The rapid, broad-spectrumefficacy makes the compositions useful as skin preparations as describedand tested in ASTM E 1173-01 provides “Standard Test Method forEvaluation of Preoperative, Precatheterization, or Preinjection SkinPreparations” and FDA Tentative Final Monograph for HealthcareAntiseptic Drug Products (TFM) (Federal Register 59 [116], Jun. 17,1994: pp. 31402-31452), which is incorporated by reference herein.

In one or more embodiments, the present invention provides adisinfectant composition that meets the standards of one or more ofEN1040 for basic bactericidal activity, EN1275 for basic fungicidalactivity, EN1500 for activity of products for use as a hygienic handrub, EN 14348 for tuberculoidal activity, EN 14476 for virucidalactivity, and EN12791 for surgical hand disinfection.

Thus, the present invention further provides a method for killing orinactivating organisms on a surface comprising applying, to the surface,an effective amount of a disinfectant composition as described herein.The disinfectant composition may be employed on a wide variety ofsurfaces or substrates, including hard surfaces, soft surfaces, skin,porous, and non-porous surfaces.

In one or more embodiments, the method provides a log reduction againstspores of at least about 1 in less than about 1 minute. In otherembodiments, the method provides a log reduction against spores of atleast about 1.5 in less than about 1 minute. In yet other embodiments,the method provides a log reduction against spores of at least about 2in less than about 1 minute. In other embodiments, the method provides alog reduction against spores of at least about 2.5 in less than about 1minute. In yet other embodiments, the method provides a log reductionagainst spores of at least about 3 in less than about 1 minute.

In one or more embodiments, the method provides a log reduction againstspores of at least about 1 in less than about 30 seconds. In otherembodiments, the method provides a log reduction against spores of atleast about 1.5 in less than about 30 seconds. In yet other embodiments,the method provides a log reduction against spores of at least about 2in less than about 30 seconds. In other embodiments, the method providesa log reduction against spores of at least about 2.5 in less than about30 seconds. In yet other embodiments, the method provides a logreduction against spores of at least about 3 in less than about 30seconds.

In one or more embodiments, the method provides a log reduction againstspores on inanimate, hard, non-porous surfaces of at least about 6 in 10minutes or less. In other embodiments, the method provides a logreduction against spores of at least about 6 in about 7 minutes or less.In yet other embodiments, the method provides a log reduction againstspores of at least about 6 in about 6 minutes or less. In otherembodiments, the method provides a log reduction against C. difficilespores on inanimate, hard, non-porous surfaces of at least about 6 inabout 10 minutes or less. In yet other embodiments, the method providesa log reduction against C. difficile spores on inanimate, hard,non-porous surfaces of at least about 6 in about 7 minutes or less. Inyet other embodiments, the method provides a log reduction against C.difficile spores on inanimate, hard, non-porous surfaces of at leastabout 6 in about 6 minutes or less.

In one or more embodiments, the method provides a log reduction againstC. difficile spores of at least about 1 in less than about 1 minute. Inother embodiments, the method provides a log reduction against C.difficile spores of at least about 1.5 in less than about 1 minute. Inyet other embodiments, the method provides a log reduction against C.difficile spores of at least about 2 in less than about 1 minute. Inother embodiments, the method provides a log reduction against C.difficile spores of at least about 2.5 in less than about 1 minute. Inyet other embodiments, the method provides a log reduction against C.difficile spores of at least about 3 in less than about 1 minute.

In one or more embodiments, the method provides a log reduction againstC. difficile spores of at least about 1 in less than about 30 seconds.In other embodiments, the method provides a log reduction against C.difficile spores of at least about 1.5 in less than about 30 seconds. Inyet other embodiments, the method provides a log reduction against C.difficile spores of at least about 2 in less than about 30 seconds. Inother embodiments, the method provides a log reduction against C.difficile spores of at least about 2.5 in less than about 30 seconds. Inyet other embodiments, the method provides a log reduction against C.difficile spores of at least about 3 in less than about 30 seconds.

Advantageously, the rapid efficacy of the methods and compositions ofthe present invention make it feasible to utilize them for skin/handdisinfection.

In one or more embodiments, the method provides a log reduction againstC. difficile spores on skin of at least about 1 in less than about 1minute. In other embodiments, the method provides a log reductionagainst C. difficile spores on skin of at least about 1.5 in less thanabout 1 minute. In yet other embodiments, the method provides a logreduction of C. difficile spores on skin of at least about 2 in lessthan about 1 minute. In other embodiments, the method provides a logreduction against C. difficile spores on skin of at least about 2.5 inless than about 1 minute. In yet other embodiments, the method providesa log reduction against C. difficile spores on skin of at least about 3in less than about 1 minute.

In one or more embodiments, the method provides a log reduction againstC. difficile spores on skin of at least about 1 in less than about 30seconds. In other embodiments, the method provides a log reductionagainst C. difficile spores on skin of at least about 1.5 in less thanabout 30 seconds. In yet other embodiments, the method provides a logreduction against C. difficile spores on skin of at least about 2 inless than about 30 seconds. In other embodiments, the method provides alog reduction against C. difficile spores on skin of at least about 2.5in less than about 30 seconds. In yet other embodiments, the methodprovides a log reduction against C. difficile spores on skin of at leastabout 3 in less than about 30 seconds.

The methods of the present invention include the step of applying adisinfectant composition to a surface.

Advantageously, good efficacy is achieved by the methods of the presentinvention when the disinfectant composition is applied to the surface atstandard temperature and at close to standard pressure. In one or moreembodiments, the temperature of the disinfectant composition whenapplied to the surface may be less than about 150° F., in otherembodiments, less than about 120° F., and in other embodiments, lessthan about 105° F. In one or more embodiments, the temperature of thedisinfectant composition may be in the range of from about 40° F. toabout 150° F., in other embodiments in the range of from about 40° F. toabout 105° F., and in other embodiments, in the range of about 70° F. to105° F.

Although the liquid disinfectant compositions of the present inventionmay be applied to the surface to be cleaned by spraying, no highpressure application is required. During this step, the disinfectantcomposition may be brought into contact with the target surface inbursts or in a continuous manner by circulating, flooding, spraying,foaming or fogging. The step may also be carried out by forming a twophase annular mist of antimicrobial treatment solution and air.

Advantageously, the methods of the present invention provide goodefficacy against spores within 10 minutes or less. Embodiments of theinvention provide good efficacy against spores within 7 minutes or less.Embodiments of the invention provide good efficacy against spores within6 minutes or less. Embodiments of the invention provide good efficacyagainst spores within 5 minutes or less. Embodiments of the inventionprovide good efficacy against spores within 2 minutes or less.Embodiments of the invention provide good efficacy against spores within1 minute or less. Embodiments of the invention provide good efficacyagainst spores within 30 seconds or less. Advantageously, the methods ofthe present invention provide good efficacy against C. difficile sporeswithin 5 minutes or less. Embodiments of the invention provide goodefficacy against C. difficile spores within 2 minutes or less.

In one or more embodiments, the method provides a log reduction againstspores, such as C. difficile, of at least about 1 in less than about 1minute. In other embodiments, the method provides a log reductionagainst spores of at least about 1.5 in less than about 1 minute. In yetother embodiments, the method provides a log reduction against spores ofat least about 2 in less than about 1 minute. In other embodiments, themethod provides a log reduction against spores of at least about 2.5 inless than about 1 minute. In yet other embodiments, the method providesa log reduction against spores of at least about 3 in less than about 1minute.

In one or more embodiments, the method provides a log reduction againstspores of at least about 1 in less than about 30 seconds. In otherembodiments, the method provides a log reduction against spores of atleast about 1.5 in less than about 30 seconds. In yet other embodiments,the method provides a log reduction against spores of at least about 2in less than about 30 seconds. In other embodiments, the method providesa log reduction against spores of at least about 2.5 in less than about30 seconds. In yet other embodiments, the method provides a logreduction against spores of at least about 3 in less than about 30seconds.

Advantageously, the methods of the present invention provide goodefficacy against C. difficile spores within 5 minutes or less.Embodiments of the invention provide good efficacy against C. difficilespores within 2 minutes or less. Embodiments of the invention providegood efficacy against C. difficile spores within 1 minute or less.Embodiments of the invention provide good efficacy against C. difficilespores within 30 seconds or less. Thus, in one or more embodiments, theduration of contact of the disinfectant composition with the targetsurface is from about 20 seconds to 5 minutes, in other embodiments,from about 25 seconds to about 2 minutes, and in other embodiments, fromabout 30 seconds to about 1 minute. It will be understood that, in someembodiments, a longer contact time is advantageous, and in one or moreembodiments, the contact time may be up to 30 minutes, and in otherembodiments, up to about 60 minutes.

The amount of disinfectant composition to be applied to the targetsurface is not particularly limited. At a minimum, a sufficient amountof disinfectant composition should be applied to substantially wet thesurface such that the surface will remain wet for the desired contacttime, noting that there will be some evaporation of the disinfectantcomposition.

Any amount of the disinfectant composition may be used for eachapplication, so long as it is at least an effective amount to contactsubstantially the entire target surface and keep it wet for at least 30to 60 seconds. In one or more embodiments, the amount of thedisinfectant composition is sufficient to contact substantially theentire target surface and keep it wet for a contact time of at least 5minutes. In one or more embodiments, the amount of the disinfectantcomposition is sufficient to contact substantially the entire targetsurface and keep it wet for a contact time of at least 6 minutes. In oneor more embodiments, the amount of the disinfectant composition issufficient to contact substantially the entire target surface and keepit wet for a contact time of at least 7 minutes. In one or moreembodiments, the amount of the disinfectant composition is sufficient tocontact substantially the entire target surface and keep it wet for acontact time of at least 10 minutes. In one or more embodiments, theamount of the disinfectant composition is sufficient to contactsubstantially the entire target surface and keep it wet for a contacttime of at least 30 minutes. In one or more embodiments, the amount ofthe disinfectant composition is sufficient to contact substantially theentire target surface and keep it wet for at least 60 minutes.

Advantageously, to minimize any possible instability or degradation ofthe active components of the compositions of the present invention, thecompositions of the present invention may be generated at the time ofuse.

In one or more embodiments, the sporicidal disinfectant composition maybe prepared by combining two or more liquid pre-mix compositions. Afirst pre-mix composition may comprise a concentrate of the peracid, asecond pre-mix composition may comprise a concentrate of the alcohol,and the acidifying agent may be present in either or both pre-mixes,such that combination of the pre-mix compositions results in adisinfectant composition comprising alcohol and a peracid at low pH asdescribed hereinabove.

In other embodiments, a first pre-mix composition may comprise aconcentrate of the alcohol and peracid, the second pre-mix compositionmay comprise a diluent, and the acidifying agent may be present ineither or both pre-mixes, such that combination of the pre-mixcompositions results in a disinfectant composition comprising alcoholand a peracid at the concentrations and pH as described hereinabove.

In one or more embodiments, the pre-mix compositions may be stored in adual dispenser and mixed only when the dispenser is actuated. Thepre-mix components may be dispensed from physically separate packages orfrom a unitary package having non-communicating chambers. For purposesof this specification, the term dual dispenser apparatus refers to aconfiguration where multiple liquid pre-mix components are dispensedfrom a plurality of physically separate packages, and also refers to aconfiguration where multiple liquid pre-mix components are dispensedfrom a unitary package having a plurality of non-communicating chambers,each chamber having an orifice through which an aliquot of a componentis dispensed.

In one or more embodiments, aliquots of the pre-mix components aredispensed substantially simultaneously, such that the liquid aliquotsare commingled. In particular embodiments, the aliquots are dispensedthrough orifices that are configured to enable the commingling of thealiquots. It will be understood that the dispenser may take a variety offorms, and may include a variety of components and configurations inorder to cause the desired commingling of aliquots of the pre-mixcomponents and dispensing of a product.

One embodiment of an exemplary dispenser is shown in FIG. 1 and isgenerally indicated by the numeral 100. Dispenser 100 may include afirst reservoir 102 containing a first liquid pre-mix component (e.g.concentrated peracid pre-mix component), and a second reservoir 104containing a second liquid pre-mix component (e.g. alcoholic diluentpre-mix component). The pH-adjusting agent may be present in either orboth of the pre-mix components. As will be apparent to those skilled inthe art, and as indicated above, the first and second reservoirs 102 and104 are not in direct communication with one another, and the first andsecond pre-mix components are therefore stored separately within thedispenser. Although separate reservoirs are shown in FIG. 1, it iscontemplated that the first and second reservoirs 102 and 104 may beprovided as physically separate chambers in a single package. Each ofthe first and second reservoirs 102 and 104 is impervious to fluidtransfer therethrough, except through inlet passages 106 and 108,respectively.

Generally, the present invention provides a method of preparing adisinfectant composition, the method comprising the steps of providing adispenser having a first reservoir containing a first liquid pre-mixthat is a concentrated peracid pre-mix, a second reservoir containing asecond liquid pre-mix that is an alcoholic pre-mix, wherein thedispenser is adapted to dispense an aliquot of the first pre-mix and analiquot of the second pre-mix, such that the aliquots commingle. Uponcommingling, the aliquots of the first pre-mix and second pre-mix form adisinfectant composition comprising an alcohol and a peracid at a pH ofabout 5 or less.

In certain embodiments, a first pump 110 may be in fluid communicationwith the first reservoir 102 through the inlet passage 106, and a secondpump 112 may be in fluid communication with the second reservoir 104through the inlet passage 108. First and second pumps 110 and 112 may beany type of pump known to those skilled in the art and suitable forconveying the first and second liquid pre-mix components from the firstand second reservoirs 102 and 104. In one or more embodiments, the pumps110 and 112 may both be positive displacement pumps. The first andsecond pumps 110 and 112 discharge the first and second pre-mixcomponents through outlet passages 114 and 116, respectively. In certainembodiments, the output or displacement of the first and second pumps110 and 112 may be adjustable to vary the rate of fluid flowtherethrough. While the exemplary dispenser 100 shown and describedincludes first and second pumps 110 and 112, it is also contemplatedthat a single pump may be used, and may be in fluid communication withboth the first and second reservoirs 102 and 104.

The outlet passages 114 and 116 may each extend to a mixing nozzle 118where the first and second pre-mix components are commingled to form adisinfectant composition. The features and dimensions of the mixingnozzle 118 may be adjusted to vary the volume of each pre-mix aliquot,as well as the rate of mixing and commingling of the first and secondpre-mix components. The mixing nozzle 118 includes a dispensing passage120 through which the disinfectant composition is dispensed.

In certain embodiments, the first and second pumps 110 and 112 may beadjusted to produce substantially the same flow rate of the first andsecond pre-mix components therethrough. In other embodiments, the pumps110 and 112 may be adjusted to provide different flow rates, and incertain embodiments, the pre-mix components may be dispensedsequentially.

In certain embodiments, the first and second pumps 110 and 112 may beadjusted to select substantially the same aliquot volume for the firstand second pre-mix components. In other embodiments, the pumps 110 and112 may be adjusted to provide different aliquot volumes.

In one or more embodiments, the first and second pumps 110 and 112 maybe adapted to dispense a single dose of composition upon actuation. Inthe same or other embodiments, the first and second pumps 110 and 112may be adapted to produce a continuous flow of the disinfectantcomposition upon actuation.

In one or more embodiments, the first pre-mix may include a concentratedform of the compositions of the present invention, and the secondpre-mix may include a diluent, such that upon being dispensed, thecombination forms a composition that includes the amounts of componentstaught herein.

Thus, the present invention provides a method of dispensing anefficacious disinfectant composition, including the steps of providing adual dispenser apparatus, where the dispenser apparatus is configured todispense multiple liquid pre-mix components from a plurality ofphysically separate packages, or where the dispenser apparatus isconfigured to dispense multiple liquid pre-mix components from a unitarypackage having a plurality of non-communicating chambers, each chamberhaving an orifice through which an aliquot of a component is dispensed;said dispenser containing a first liquid pre-mix composition thatincludes a C₁₋₆ alcohol and a peracid, said first liquid pre-mixcomposition having a pH of less than about 5; said dispenser furthercontaining a second liquid pre-mix composition that includes a diluentselected from a C₁₋₆ alcohol, water, and mixtures thereof; and actuatingsaid dispenser to commingle an aliquot of said first pre-mix with analiquot of said second pre-mix and dispense said commingled aliquots.

In one or more embodiments, aliquots of the pre-mix components aredispensed substantially simultaneously, such that the liquid aliquotsare commingled.

Advantageously, compositions of the present invention exhibit improvedstability under normal conditions of product storage and transport, andtherefore exhibit longer shelf life than peroxide-based products andeven other peracid-based products that do not contain the synergisticcombination of peracid and alcohol at a low pH. Optionally, thecompositions of the present invention may be further stabilized by theinclusion of one or more stabilizers. Examples of stabilizers includechelators, inorganic acids and organic acids.

Decomposition of the peracid to peroxide leads to a reduction inefficacy. The peroxide is in turn susceptible to decomposition to water,which leads to a further reduction in efficacy. Reference to stability,within the context of the present invention, means that the peracid doesnot decompose within the disinfectant composition to peroxide or waterduring a stated period of time, and when kept within stated conditionsof temperature and pressure. Stability can be assessed by directchemical measurement of the concentration of the peracid, for example byHPLC, and/or can be assessed by measuring the efficacy of thedisinfectant composition.

In one or more embodiments, compositions of the present invention arestable, under conditions of standard temperature and pressure for atleast 3 days, in other embodiments, for at least 1 week, in otherembodiments, for at least 2 weeks, in other embodiments, for at least 1month, in other embodiments, for at least 2 months, in otherembodiments, for at least 6 months, in other embodiments, for at least 1year. Thus, compositions of the present invention are able to maintaintheir broad efficacy, including efficacy against viruses, bacterial andfungal spores such as C. difficile spores, over longer periods of time,whereas other products would have undergone decomposition and would nolonger be as efficacious.

Accordingly, the present invention provides a method of improving thestability of peracid-containing disinfectant compositions, the methodincluding the steps of combining a peracid with a C₁₋₆ alcohol, at a pHof less than about 5, to form a stabilized peracid-containingcomposition, and maintaining the composition in a container or dispenseruntil ready for use, wherein the stability of the stabilized compositionis improved when compared to the stability of the same composition butnot containing a C₁₋₆ alcohol, and not at a pH of less than about 5.

In order to demonstrate the practice of the present invention, thefollowing examples have been prepared and tested. The examples shouldnot, however, be viewed as limiting the scope of the invention. Theclaims will serve to define the invention.

EXAMPLES

In the following examples, where peroxypyruvic acid was employed, it wasobtained from CHD Bioscience under the tradename VERIOX™. VERIOX™ is aconcentrated solution of peroxypyruvic acid in water, and was diluted toachieve the concentrations described below.

Example 1 was a solution of 845 ppm peroxypyruvic acid (PPA) in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 2 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 3 was a solution of 1690 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 4 was a solution of 70 wt. % ethanol and 1690 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 5 was a solution of 6760 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 6 was a solution of 70 wt. % ethanol and 6760 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 7 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 2.

Example 8 was a solution of 70 wt. % ethanol and 8455 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about2.

Example 9 was a solution of 1690 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 2.

Example 10 was a solution of 70 wt. % ethanol and 1690 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about2.

Example 11 was a solution of 6760 PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 2.

Example 12 was a solution of 70 wt. % ethanol and 6760 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about2.

Example 13 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 2.5.

Example 14 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about2.5.

Example 15 was a solution of 1690 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 2.5.

Example 16 was a solution of 70 wt. % ethanol and 1690 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about2.5.

Example 17 was a solution of 6760 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 2.5.

Example 18 was a solution of 70 wt. % ethanol and 6790 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about2.5.

The samples were tested for efficacy against C. difficile spores,according to the ASTM E 2315 method, “Standard Guide for Assessment ofAntimicrobial Activity Using a Time-Kill Procedure.” Contact time was 30seconds. Results are shown in Table 1 and FIG. 2.

TABLE 1 Log Reduction Example Composition pH (CFU/ml) 1  845 ppm PPA 1.50.5 2  845 ppm PPA + 70% ethanol 1.5 2.44 3 1690 ppm PPA 1.5 1.02 4 1690ppm PPA + 70% ethanol 1.5 3.56 5 6760 ppm PPA 1.5 3.86 6 6760 ppm PPA +70% ethanol 1.5 3.86 7  845 ppm PPA 2 0.54 8  845 ppm PPA + 70% ethanol2 0.06 9 1690 ppm PPA 2 1.32 10 1690 ppm PPA + 70% ethanol 2 0.91 116760 ppm PPA 2 3.86 12 6760 ppm PPA + 70% ethanol 2 3.71 13  845 ppm PPA2.5 0.38 14  845 ppm PPA + 70% ethanol 2.5 −0.07 15 1690 ppm PPA 2.50.61 16 1690 ppm PPA + 70% ethanol 2.5 0.02 17 6760 ppm PPA 2.5 3.86 186760 ppm PPA + 70% ethanol 2.5 2.26

Example 19 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 20 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 21 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 22 was a solution of 1690 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 23 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 24 was a solution of 70 wt. % ethanol and 1690 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 25 was a solution of 3380 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 26 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 27 was a solution of 70 wt. % ethanol and 3380 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 28 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 29 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 30 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.8.

Example 31 was a solution of 1690 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 32 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 33 was a solution of 70 wt. % ethanol and 1690 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.8.

Example 34 was a solution of 3380 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 35 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 36 was a solution of 70 wt. % ethanol and 3380 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.8.

As for Examples 1-18, the samples were tested for efficacy against C.difficile spores, according to the ASTM E 2315 method, “Standard Guidefor Assessment of Antimicrobial Activity Using a Time-Kill Procedure.”Contact time was 30 seconds. Results are shown in Table 2 and FIG. 3.

TABLE 2 Log Reduction Example Composition pH (CFU/ml) 19  845 ppm PPA1.5 1.5  20 70% ethanol 1.5 0.3  21  845 ppm PPA + 70% ethanol 1.5 1.7722 1690 ppm PPA 1.5 2.18 23 70% ethanol 1.5 0.62 24 1690 ppm PPA + 70%ethanol 1.5 3.56 25 3380 ppm PPA 1.5 3.86 26 70% ethanol 1.5 0.43 273380 ppm PPA + 70% ethanol 1.5 4.01 28  845 ppm PPA 1.8 1.67 29 70%ethanol 1.8 0.44 30  845 ppm PPA + 70% ethanol 1.8 0.25 31 1690 ppm PPA1.8 2.16 32 70% ethanol 1.8 0.16 33 1690 ppm PPA + 70% ethanol 1.8 0.9334 3380 ppm PPA 1.8 3.41 35 70% ethanol 1.8 0.06 36 3380 ppm PPA + 70%ethanol 1.8 3.32

Example 37 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 38 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 39 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 40 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 41 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 42 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 43 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 44 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 45 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 46 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 47 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 48 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.8.

Example 49 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 50 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 51 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.8.

Example 52 was a solution of 70 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 53 was a solution of 845 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.8.

Example 54 was a solution of 70 wt. % ethanol and 845 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.8.

As for Examples 1-18, the samples were tested for efficacy against C.difficile spores, according to the ASTM E 2315 method, “Standard Guidefor Assessment of Antimicrobial Activity Using a Time-Kill Procedure.”Contact time was 30 seconds for examples 37-39 and 46-48, 60 seconds forexamples 40-42 and 49-51, and 5 minutes for examples 43-45 and 52-54.Results are shown in Table 3 and FIG. 4.

TABLE 3 Exposure Log Reduction Example Composition pH Time (CFU/ml) 3770% ethanol 1.5 30 sec 0.17 38 845 ppm PPA 1.5 30 sec 0.39 39 845 ppmPPA + 70% ethanol 1.5 30 sec 0.91 40 70% ethanol 1.5 60 sec 2.17 41 845ppm PPA 1.5 60 sec 0.45 42 845 ppm PPA + 70% ethanol 1.5 60 sec 3.36 4370% ethanol 1.5  5 min 3.17 44 845 ppm PPA 1.5  5 min 3.75 45 845 ppmPPA + 70% ethanol 1.5  5 min 3.75 46 70% ethanol 1.8 30 sec −0.21 47 845ppm PPA 1.8 30 sec 0.57 48 845 ppm PPA + 70% ethanol 1.8 30 sec −0.05 4970% ethanol 1.8 60 sec 0.36 50 845 ppm PPA 1.8 60 sec 0.58 51 845 ppmPPA + 70% ethanol 1.8 60 sec 2.00 52 70% ethanol 1.8  5 min 3.60 53 845ppm PPA 1.8  5 min 3.75 54 845 ppm PPA + 70% ethanol 1.8  5 min 3.75

Example 55 was a solution of 500 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 56 was a solution of 60 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 57 was a solution of 60 wt. % ethanol and 500 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 58 was a solution of 500 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 59 was a solution of 80 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 60 was a solution of 80 wt. % ethanol and 500 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 61 was a solution of 1000 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 62 was a solution of 60 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 63 was a solution of 60 wt. % ethanol and 1000 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

Example 64 was a solution of 1000 ppm PPA in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 65 was a solution of 80 wt. % ethanol in water, to which 12 Nhydrochloric acid had been added to achieve a pH of about 1.5.

Example 66 was a solution of 80 wt. % ethanol and 1000 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5.

As for Examples 1-18, the samples were tested for efficacy against C.difficile spores, according to the ASTM E 2315 method, “Standard Guidefor Assessment of Antimicrobial Activity Using a Time-Kill Procedure.”Contact time was 30 seconds. Results are shown in Table 4 and FIG. 5.

TABLE 4 Log Reduction Example Composition pH (CFU/ml) 55  500 ppm PPA1.5 0.51 56 60% ethanol 1.5 0.56 57  500 ppm PPA + 60% ethanol 1.5 1.5458  500 ppm PPA 1.5 0.51 59 80% ethanol 1.5 1.20 60  500 ppm PPA + 80%ethanol 1.5 3.70 61 1000 ppm PPA 1.5 0.50 62 60% ethanol 1.5 0.56 631000 ppm PPA + 60% ethanol 1.5 1.62 64 1000 ppm PPA 1.5 0.50 65 80%ethanol 1.5 1.20 66 1000 ppm PPA + 80% ethanol 1.5 2.13

Example 67 was just a bland handwash, and the contact time was 60seconds.

Example 68 was a solution of 3000 ppm PPA in water. The pH was about 2-3and the contact time was 30 seconds.

Example 69 was a solution of 70 wt. % ethanol and 3000 ppm PPA in water,to which 12 N hydrochloric acid had been added to achieve a pH of about1.5, and the contact time was 30 seconds.

Example 70 was a solution of 8000 ppm PPA in water, the pH was about2-3, and the contact time was 30 seconds.

The samples were evaluated for efficacy against C. difficile sporesaccording to the following in vivo test protocol. (Room temperature).

In Vivo C. difficile Spore Test Method Protocol:

Spore Growth Media and Neutralizer

-   -   BPB+: Butterfield's Phosphate Buffer    -   BHIT-AMP: Brain Heart Infusion Agar with Sodium Taurocholate        hydrate (0.1%) and Ampicillin (0.2 μg/ml)

Inoculum

-   -   Purified C. difficile spores (ATCC 700057) at an approximate        concentration of 8.5 log₁₀ CFU/ml. Spores were suspended in        sterile water and stored at −80° C., and allowed to come to room        temperature prior to testing.

Pretreatment of Hands

-   -   Pre-wash hands with bland soap (GOJO® Clear & Mild Foam        Handwash) and pat dry with paper towels and wait five minutes        before applying spore inoculum to hands

Inoculation of Fingertips with C. difficile Spores

-   -   Dispense 5 μL of the spore suspension directly onto the index,        middle, and ring fingerpads of each hand    -   Rub opposite fingerpads together on and off until dry    -   Wait at least 1 minute before determining the baseline        contamination of the fingers

Baseline Recovery

-   -   Sample one finger on each hand separately in standard size petri        dishes (100×15 mm) containing 5 mL BPB+ by rubbing for one        minute    -   Pat fingerpads dry on paper towels to remove excess BPB+

Product Application

-   -   For Handwash: briefly wet hands, apply product (2 pumps of        foaming hand washes), lather for 60 seconds, rinse off excess        lather for 10 seconds, lightly pat dry    -   For disinfectant compositions: (Examples): rub fingertips        together under a continuous stream from a wash bottle filled        with the appropriate product for the desired exposure time,        briefly rinse to neutralize, lightly pat dry

Post Product Exposure Recovery

-   -   Sample index, middle, and ring fingers of each hand together as        described above

Enumeration of C. difficile

-   -   Dilute baseline and post product exposure recovery in BPB+ and        enumerate on BHIT-AMP.

Log Reduction Calculation

-   -   After enumeration of viable C. difficile with subtract post        exposure recovery from baselines to get a log reduction

Example 71—A solution of 4000 ppm peroxypyruvic acid (PPA) in water wasprepared, using distilled, deionized water. The solution was maintainedin a vented container at 22° C. The concentration of PPA was monitoredover time by using HPLC. After about 24 hours, the concentration of PPAhad dropped to about 0, indicating that the composition was not stableover that period of time.

Various modifications and alterations that do not depart from the scopeand spirit of this invention will become apparent to those skilled inthe art. This invention is not to be duly limited to the illustrativeembodiments set forth herein.

What is claimed is:
 1. A composition for inactivation or kill of bacterial and fungal spores, the composition comprising: a C₁₋₆ alcohol, and a peracid, wherein the disinfectant composition is characterized by a pH of no more than about 5, and wherein the composition exhibits a synergistically enhanced efficacy against C. difficile spores, when compared to the efficacy of the alcohol or peracid alone.
 2. The composition of claim 1, wherein the disinfectant composition comprises at least about 30 wt. % of the C₁₋₆ alcohol, based upon the total weight of the disinfectant composition.
 3. The composition of claim 2, wherein the peracid is present in an amount of from about 100 ppm to about 2 wt. %, based upon the total weight of the disinfectant composition.
 4. The composition of claim 3, wherein the peracid is peroxypyruvic acid.
 5. The composition of claim 4, where tine pH of the composition is from about 0.5 to about
 3. 6. The composition of claim 1, wherein the peracid is selected from the group consisting of C₁-C₁₀ percarboxylic acids, diperoxyglutaric acid, diperoxyadipic acid, diperoxysuccinic acid, diperoxysuberic acid, diperoxymalonic acid, peroxylactic acid, peroxyglycolic acid, peroxyoxalic acid, peroxypyruvic acid, and mixtures thereof.
 7. The composition of claim 1, wherein the peracid is present in an amount of from about 100 ppm to about 2 wt. %, based upon the total weight of the disinfectant composition.
 8. The composition of claim 1, wherein the pH of the composition is from about 0.5 to about
 3. 9. A method for disinfection of a surface, the method comprising: contacting the surface with the composition of claim
 1. 10. The method of claim 9, wherein the pH of the composition is from about 0.5 to about
 3. 11. The method of claim 9, wherein at least 1 log₁₀ reduction is achieved against C. difficile spores within about 5 minutes, under conditions of standard temperature and pressure.
 12. The method of claim 9, wherein at least 1 log₁₀ reduction is achieved against C. difficile spores within about 1 minutes, under conditions of standard temperature and pressure.
 13. The method of claim 9, wherein at least 1 log₁₀ reduction is achieved against C. difficile spores within about 30 seconds, under conditions of standard temperature and pressure.
 14. The method of claim 9, where said log reduction is achieved on skin. 